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Mission Folder: View Mission for 'Silver Bullet'

State Texas

Grade 7th

Mission Challenge Food, Health and Fitness

Method Scientific Inquiry using Scientific Practices

Students anthrobones dinosaur24 silvershade15 skittles02

Team Collaboration(1) Describe the plan your team used to complete your Mission Folder. Be sure to explain the role of each team member and how you shared and assigned responsibilities. Describe your team’s process to ensure that assignments were completed on time and deadlines were met.

The United States is one of the wealthiest nations on Earth, yet serious health issues still affect many Americans. A crisis is mounting as the number of people diagnosed with diabetes skyrockets. More than 29 million people are infected with this disease, and developing a chronic wound that refuses to heal is an expected complication. Diabetics are particularly in fear of wounds because if they wait too long, it can get to the point of no return and eventual amputation. This is just one of the very scary and common problems for diabetics across the country. As middle school students, we studied the facts and as if that was not reason enough to work on the problem, each of us knows someone personally who suffers from diabetes and who has experienced issues with wounds. For these reasons, the team selected this challenging national issue.

Silver Bullet is a team of seventh graders that began work in April of 2015. We began studying a substance known as bacterial biofilms that are the root of the problem in chronic wounds. This Ecybermission project seeks to build awareness of biofilms, infections, and contribute to advancing a way to fight the problem.

Our team is involved in many different school activities such as football, basketball, science/math competitions, music, and theatre. We had to find time in our schedules when everyone could meet. Our school has set aside time for extra help on Wednesday afternoons and we would meet then to share research, conduct experiments, and work with community partners on the project. We also met on Sunday afternoons in the school science lab for experimentation and after school as needed so that deadlines were always met. Several field trips were taken during school because the principal is supportive of STEM. Everybody was very determined to find an answer to the problem of chronic wounds.

The plan to complete the project was to meet regularly, assign tasks and hold ourselves to deadlines, search for experts who would allow us to collaborate with them and use their labs, and agree to fight procrastination. The team completed the Mission Folder answers by dividing the questions into groups, and assigning the questions as evenly as possible to each person, while keeping in mind the strengths and weaknesses of the team members. When there were conflicts about what to do next or how to read the data, the team talked through all possible answers and then came to a unanimous decision. Since team members have completed projects before, working together and maximizing time was not difficult.

Each team member worked hard on each aspect of the project. We followed our action plan, kept a binder in the classroom that kept everyone on track, and each person helped the others stay focused. Other Ecybermission teams at our school did not finish, but it was great to be part of this team!

To encourage each other this year, we kept a positive attitude during team meetings and never allowed put-downs of anybody or any idea. We also complimented each other on ideas, even if we personally thought there was a better way to do something. Together, we celebrated successes along the way, texted each other, and talked about the project all year. In all these ways, team members encouraged one another to finish the project.

During team meetings, a deadline was assigned for all questions to be completed and turned in to our Team Advisor so it could be read by an adult. Suggestions were made for editing and then the Team Advisor submitted the final copy of the Mission Folder with the uploaded files as evidence of the work we accomplished this year. The experience of being part of Ecybermission and working towards a common goal was exciting and allowed many opportunities to work in university laboratories and meet with doctors that became role models.

Elizabeth fulfilled the role of leader, organizer, writer, and artist. She scheduled meetings and kept the team informed of upcoming events and deadlines.

Kaden's roles included data analyzer, creative thinker, troubleshooting, research online, and presentations.

Blade was the dreamer, public speaker, team motivator, website builder, and took charge during in vitro and in vivo testing.

Kaylah was in charge of interviews, public speaking, engineering hydrogels with silver, and research.

A key factor in the ability to complete the mission was teamwork.

Please open the table showing the specific roles and responsibilities of each member, the timeline for the project, and an overall view of the project in the uploaded pdf files titled “Team Responsibilities,” “Action Plan,” and “Timeline.”

of 9eCYBERMISSION - Advisor Section

4/21/2016https://www.ecybermission.com/Advisor/ViewMissionFolder_ReadOnly_SM/13253

Uploaded Files:

• [ View ] Team Responsibilities (By: dinosaur24, 02/24/2016, .pdf)

A chart was created to show the roles and responsibilities each of us fulfilled during this project.

• [ View ] Action Plan (By: anthrobones, 02/27/2016, .pdf)

A graphic organizer and flow chart was created to show how our team went from a community problem, to a series of hypotheses, to a

set of experiments, and finally to community solutions to the problem of chronic wound care.

• [ View ] Timeline (By: anthrobones, 02/27/2016, .pdf)

A timeline of the project is included to demonstrate progress from April 2015 - May 2016. Even though the competition deadline is

February 29, there are exciting events scheduled throughout the spring.

Scientific Inquiry

Problem Statement

(1) What problem in your community did your team try to solve? Why is this problem important to your community?

Oil field workers, welders, farmers, and ranchers – these are the working people of our community. According to Covenant Hospital, they are also the people who suffer from multiple injuries and oftentimes have wound issues. While most wounds are relatively harmless and heal quickly, an unattended wound on an individual with compromised health can become a serious infection.

Chronic wounds are rarely seen in individuals who are healthy. In fact, most people suffering from chronic wound complications are affected with other diseases such as obesity or diabetes. The United States National Institutes of Health states that chronic wounds affect around 6.5 billion people. For diabetics, the leading health concern in this state, the unfortunate solution is oftentimes amputation or death.

According to Centers for Disease Control and Prevention, over 29 million Americans suffer from diabetic wounds over the years. This shocking statistic was the motivation for selecting this community problem. In a county with a high rate of poverty, a large elderly population, and diabetes, finding a solution for healing wounds would have a lasting change on this community and many others. The National Institutes of Health states that $25 billion is spent to treat life-threatening wounds. It goes without saying that even though healing wounds is expensive, having a chronic wound healed is worth every penny.

Problem Statement: Can an affordable and effective material and bandage be developed to aid in the control and treatment of diabetic chronic wounds?

(2) List at least 10 resources you used to complete your research (e.g., websites, professional journals, periodicals, subject matter experts).

Chronic wounds are an issue in this community and throughout the nation. The team began research by identifying subject matter experts, community leaders, and organizations near our town that would provide a strong foundation for this project. The information came from webinars, TED talks, newspapers, online podcasts, interviews, professional journals, emails, websites, and newspapers. Government agencies and independent studies were trusted research sites. The hypotheses and experiments are based on what was learned from these sources in hopes for a solution that will benefit many individuals. Please see three uploaded files “Bibliography,” “Collaboration with Community Experts,” and “Contact List” for evidence of thorough and valid research.

Bibliography:

Anthes, E. 7 Next-Gen Bandages That Help Heal Wounds. Retrieved August 20, 2015.

Auddy, Runa. "New Guar Biopolymer Silver Nanocomposites for Wound Healing Applications." http://www.hindawi.com/journals/bmri/2013/912458. BioMed Research International, 18 Aug. 2013. Web. 4 Oct. 2015.

Brickey, Henri. "Trials to Showcase Power of Phages in Fighting Infection." Lubbock Avalanche-Journal 4 Jan. 2007. Print.

Cardinal, Matthew, David E. Eisenbud, and David G. Armstrong. "Wound Shape Geometry Measurements Correlate to Eventual Wound Healing." Wound Repair and Regeneration 17.2 (2009): 173-78.

Cevher, E. (2011, November 16). Biopolymers as Wound Healing Materials: Challenges and New Strategies. Retrieved November 16, 2015.

Cueva, Angel, MD. “The Techniques of using Pipettes for Quantifying Research.” Personal interview conducted by Blade Henry, Kaden Moses, Kaylah Nunn, and Elizabeth Casarez. 12 Feb 2016.

Dissanaike, Sharmila, MD. "TTUHSC Surgery Burn Center of Research Excellence (BCoRE)." TTUHSC. 13 Dec. 2015.

Donlan, Rodney M. "Biofilms: Microbial Life on Surfaces." www.ncbi.nih.gov. Rodney M. Donlan, 13 Dec. 2015. Web.

Engelking, Carl. "Nanotech Lotion Speeds Diabetic Wound Healing." Discover Magazine. Kalmbach Publishing, April 2016 Issue: Subscribers Sneak Peek. Web. 10 Feb. 2016.

Fleming, Derek, MD. “The Role of a Medical Technologist.” Personal Interview conducted by Kaden Moses and Blade Henry. 12 Feb 2016.

Harada, Cesar. "How I Teach Kids to Love Science." TEDX. N.p., n.d. Web. 4 Sept. 2015.

Lehman, Michael, MD. “Antibiotic Resistance and Treating Biofilms for Success in Healing Chronic Wounds.” Personal Interview by Kaylah Nunn. 17 Jan 2015.

Martin, Normin. "Yang Hu Takes New Post with Tech’s Department of Plant and Soil Science." www.depts.ttu.edu. CASNR News Center, 13 Dec. 2015. Web. 25 Feb. 2016.

Nightingale, Sarah. "Show 'Health Care Heroes' Features Lubbock Doctors Known for Treating Hard-to-heal Wounds." Lubbock Avalanche-Journal 8 Sept. 2010. Print.

Paddock, Catharine, PhD. "MRSA / Drug Resistance Infectious Diseases / Bacteria / Viruses Anti-biofilm Molecule May Help Fight Superbugs." Medical News Today. MNT, 23 May 2014. Web. 8 Sept. 2015.

Pastore, Rose. "How A Simple New Invention Seals A Gunshot Wound In 15 Seconds." Popular Science. Bonnier Corporation Company, 03 Feb. 2014. Web. 12 July 2015.



Richards, Summer, MD. "Biofilms and Their Role in Infection." Personal interview conducted by Kaylah Deavours Nunn. 4 Feb. 2016.

Rogers, Gregory, MD. “Prevalence of Chronic Wounds in Emergency Rooms and Clinics of Rural West Texas.” Personal Interview conducted by Kaylah Nunn, Blade Henry, Elizabeth Casarez, Kaden Moses. 10 Oct 2015.

Rumbaugh, Kendra. "Department of Surgery - The Rumbaugh Lab." www.ttuhsc.edu. Kendra Rumbaugh, 13 Dec. 2015. Web. 02 Oct 2015.

Sezer, Ali. "Biopolymers as Wound Healing Materials: Challenges and New Strategies." www.intechopen.com. InTech, 16 Nov. 2011. Web. 4 Oct. 2015.

Venkatrajah, B. "Biopolymer and Bletilla Striata Herbal Extract Coated Cotton Gauze Preparation for Wound Healing." http://scialert.net PDF, 24 Apr. 2013. Web. 4 Oct. 2015.

Wolcott, Randy, MD. “Treatment Alternatives for Chronic Wounds: Fighting Biofilms.” Interview by Kaylah Nunn, Elizabeth Casarez, Kaden Moses, Blade Henry. 21 Jan 2015.

Zhao, Hannah, MD. “Preparing Petri Dishes for Bacterial Growth and Analysis.” Personal Interview conducted by Kaylah Nunn and Elizabeth Casarez. 12 Feb 2016.

Zhao, X. PhD. Stretchable Hydrogel Electronics: Water-based “Band-Aid” senses temperature, lights up, and delivers medicine to the skin.” MIT News. Retrieved December 7, 2015.

(3) Describe what you learned in your research.

Biodegradable Bandages --- Biodegradable bandages are being created that act as a new skin forming over the wound. First the doctors attach the patient to a scaffold before the dressing is applied over the wound. Then the doctors take a biopsy of the patient’s skin cells. The skin cells multiply and grow over the scaffold so when the bandage eventually dissolves, all you have left is the patient’s own skin cells. What if this technology could be combined with the hydrogel technology and wounds could be covered with medicated and moist bandages that would never need to be removed? If that happened, biofilms might not be able to get ahead.

History of Wounds --- Wound healing is an important process that scientists have been trying to make better for hundreds of years. Natural and artificial ways of wound healing exist as doctors fight biofilms and infection. For replacing tissue in chronic wound patients, scientists are turning to biocompatible and biodegradable polymers. These are alternatives to painful skin grafting, in addition to tissue engineering to speed up wound healing.

Herbal Uses --- As an accelerant of wound healing (to speed it up), polymer solutions were spread onto cotton gauze and Bletilla striata, an herb, was added to the gauze to increase wound healing. The cotton gauze was tested and showed to have rapid success when closing up the chronic wound.

Bandages --- Bandages for wounds come in many shapes and sizes but there is a new kind of bandage called a “hydrogel” that may speed up the healing process. It has several advantages that others do not such as being very flexible so it will stay in place no matter where the wound occurs. It is made of a material that is mostly water but it is also stretchy. Since the human body is wet and soft, this bandage is more similar to the needs of the wound. What if hydrogels could be injected with medications that would fight biofilms? If so, the hydrogel could be placed on the wound, it would stay in place and keep the wound moist, and the material would fight biofilms and infections they cause.

Biofilms --- Biofilms are like communities of bacteria but they are very well-organized. They stick to each other and they are microorganisms sometimes called “slime” in the medical field. Usually they are on the surface of tissues and also on surfaces in hospitals or other settings. Biofilms cause more than 65% of infections with their polysaccharide covers. The problem with biofilms comes when it allows the bacterial colony to communicate with each other and fight any kind of treatment that a doctor may be applying. Some antibiotics produce glycogen and that gives the biofilms plenty of food. Sometimes, what a person thinks is an antibiotic resistant infection that will not clear up, may really be the result of biofilms working overtime.

Resistance to Antibiotics --- Bacteria can be grouped as “gram negative” or “gram positive” and it depends on what kind of cell walls they have. Some can be stopped by certain kinds of antibiotics but according to an immunology professor at the University of British Columbia Bob Hancock, “Our entire arsenal of antibiotics is gradually losing effectiveness.” This is where the problem of chronic wounds comes in – antibiotics cannot always get rid of the infection in that wound because the biofilms are so good at keeping them out. Since many antibiotics are becoming ineffective in the fight against infection, what about trying different types of materials that the biofilms and bacteria may not have seen before?

Biomaterial-based Dressing --- Closing wounds and burns completely can be done through a method called autografting that moves skin into a new position on the body from which it was removed. However inadequate donor areas for large led to a new search for a new tissue source. Biological dressings are natural dressings with collagen-type structures.

Water Based Dressing --- Scientist Xuanhe Zhao has designed a new Water-based bandaid that will help promote healing faster. First he makes sure that the bandage will stay bonded to really flexible areas of the body such as a knee or elbow. Then Zhao gets the hydrogel which is a rubbery material mostly composed of water. Zhao then connects the conductive wires to the bandage with the semiconductor chips, and temperature sensors. Electronics are usually hard and dry but in this case the human body is soft and wet. These two systems have drastically different properties. So when you make the electronics devices you want to make them soft and stretchy to fit the environment of the human body. His team came up with a way to use hydrogels by mixing water with biopolymers to create stretchy materials with also stiffness so they will hold. Zhao also added LED lights that could attach to different sections of the body.

Hypothesis

(4) State your hypothesis. Describe how your hypothesis could help solve your problem.

From research to interviews, several substances were chosen to be tested in the development of a wound ointment and enhanced bandage. The materials were selected based on their properties and potential to fight biofilms surrounding bacteria in wounds. The following would be used in forming the hypotheses:

Colloidal silver is an antibacterial material chosen because of its astonishing ability to control antibiotic resistant superbugs in other tests. In addition, silver is antimicrobial and anti-inflammatory, therefore it would possibly be a key in healing stubborn wounds.

Xylitol was recommended by Dr. Wolcott because it is easily accessible for use in the lab. Bacteria in wounds often feed on glucose and since xylitol is a sugar alcohol, it blocks the energy-producing pathway of the bacteria - basically robbing it of food. According to the research, xylitol may eliminate harmful bacterial colonies by 27% to 75% in the mouth, therefore it would be tested in vitro for possible success in topical wounds.

Probiotics were selected because it is a well-known fact that probiotics add good bacteria to the gut and can fight off bad bacteria internally. However, after visiting an Ecybermission recommended site called LiveScience, the team learned some evidence is linked to its benefit as a topical treatment as well. A common probiotic,



fructooligosaccaride, was selected for in vitro experimentation.

The following hypotheses were written to specifically address the problem statement. They are able to be measured quantitatively and qualitatively, repeated by anyone for reliable results, and are research-based to be valid.

Hypothesis 1 - In Vitro Experiment: If a composite material of xylitol plus colloidal silver is created and is effective in preventing the spread of bacteria during in vitro experimentation, (as quantified by measuring zones of inhibition), then the substances could be useful as a topical ointment for chronic wounds.

Hypothesis 2 -In Vivo Experiment: If a material can successfully lead to cell regrowth, as evidenced by in vivo experiments by decreasing the length of incisions on invertebrates, then the substance could be used to promote healing of chronic wounds.

Hypothesis 3 – Bandage Enhancement: If colloidal silver, xylitol, silver/xylitol composite, or probiotics could be successfully applied within a bandage, then the enhanced bandage could be more effective in sealing wounds from airborne bacteria and in fighting biofilms and infection with its embedded ointment.

(5) Identify the independent variables and the dependent variables in your hypothesis.

For hypothesis 1 - In Vitro Experiment - The independent variable is the material applied to the Petri dish agar to fight bacteria. This includes colloidal silver, probiotics, xylitol, and the xylitol/silver composite. The dependent variable is the size of the zone of inhibition measured in millimeters surrounding each of the materials within the Petri dishes.

For hypothesis 2 -In Vivo Experiment – The material most effective in preventing bacterial growth during the in vitro experiment will become the independent variable in this experiment – either silver, xylitol, probiotics, or the silver/xylitol composite. The dependent variable is the size of the invertebrate’s wound as measured in millimeters over a period of time.

For hypothesis 3 – Bandage Enhancement: The independent variable is the kind of ointment applied to the bandage, and the dependent variable is the amount of ointment remaining in the bandage over a period of time.

(6) How did you measure the validity of your hypothesis? (In other words, how did you determine that your hypothesis measures what it is SUPPOSED to measure?)

In vitro hypothesis validity - If the zone of inhibition surrounding the applied materials (silver, xylitol, xylitol + silver, or probiotic) is at least 10 mm in diameter, the hypothesis will be proven valid. This would indicate a clear area free of bacterial growth as a result of the material application. Measurements will be made using millimeters from the center of a material-soaked disk outward to the nearest bacterial colony present. The disks are used to control the size of application of the material within the Petri dish for accurate and reliable measurements. According to Dr. Rumbaugh in the Texas Tech wound care laboratory, a zone at least double the size of disk application would indicate a successful treatment.

In vivo hypothesis validity – The validity of the hypothesis will be proven if the length of the incision on the earthworm decreases over time as a result of the applied material. Testing will only be conducted on earthworms and NOT on vertebrate organisms. Care will be taken to conduct the experiment in a professional and careful manner, knowing that even invertebrates are animals to be respected.

Bandage enhancement validity – The validity of the hypothesis will be supported if at least 80% of the healing material is able to be embedded in the bandage for application. If the bandage is able to keep the material as part of its structure, then the ointment would be available on the wound surface and could promote healing.

Experimental Design

(7) List the materials you used in your experiment. Include technologies you used (e.g., scientific equipment, internet resources, computer programs, multimedia, etc.).

In Vitro Experiment ---

Pipette GlovesGogglesPetri dishesSaucepan and wood spoonHotplateTriple beam balanceBeakersThermometerScoopIncubator Distilled WaterGraduated CylindersMetric measuring spoonsCoffee FiltersHole PunchAlcohol for sterilizing equipmentNutrient agarColloidal Silver – 500 ppmXylitol CrystalsFructooligosaccaride – Probiotic

In Vivo Experiment ---

Colloidal Silver – 500 ppmXylitolAlcohol for sterilizing equipmentMetric measuring spoonsGraduated CylindersDistilled WaterThermometerBeakersTriple beam balance



Earthworms, Lumbricus terrestrisScalpelForcepsMetric rulerPyranineYellow Highlighter MarkersBlack LightSyringe and needleSterile bowlsSoilGogglesGlovesPipette

Bandage Enhancement ---

HydrogelPadsSyringe and needlesDistilled waterBeakerMetric measuring spoonsSilverPyranine fluorescent chemical

Technology Used ---

Compound Light MicroscopeApple ComputerYouTube instructional videosTED X Talks – microbiologyBlack lightMac BookCell phoneIncubatorDigital CameraMicrosoft ExCelMicrosoft PowerPointMicrosoft WordGoogle SketchUpWeebly Website BuilderGoogle DocsGoogle Drive

(8) Identify the control group and the constants in your experiment.

In vitro Experiment

Control – There are often two controls used when conducting tests on bacterial growth and this experiment utilized both controls. The first control is a Petri dish containing only nutrient agar and no bacteria. The second control is a Petri dish containing nutrient agar and bacteria, but lacking a substance to control its growth. Both control dishes will be placed alongside the test samples for all three trials/tests being conducted.

Constants – Constants are the quantities in the experiment that are not allowed to be changed, either because they cannot be or because the scientist does not choose to change them. Constants can be physical (such as pi, average body temperature, boiling point of water) or can be control constants that are kept the same for the purpose of having an experiment that can be reproduced by someone else (such as holding the temperature steady, or the length of time the experiment is run, or volume of liquids tested).

The physical constant in this experiment is bringing water to a boil at 100oC for preparing the agar properly. The control constants include following the written procedures exactly for all three trials; maintaining the incubator at a temperature of 37oC to mimic body temperature for bacterial growth; using the same size disks for each material; collecting the data 16 hours after incubation for all tests; using the same amount of water and agar for preparing the Petri dishes; adding the same amount of agar to each dish; soaking the filter paper in the same amount of each material for the same amount of time; using standard-sized Petri dishes divided into equal quadrants for multiple trials; measuring the zone of inhibition from the center of each paper disk outward to the first colony of bacteria present.

In vivo Experiment

Controls – Earthworms with incisions the same length as that of the three test subjects will be used to compare the effectiveness of the colloidal silver. There will be no ointment applied to the control earthworm.

Constants – The control constants include using isopropyl alcohol to clean scalpels for exactly 45 seconds each time before making any incisions; making a 1.0 centimeter incision in each earthworm tested; measuring the length of the incision using a flexible dissecting ruler each time; measuring the control and the test subjects within the same hour; applying the same amount of silver ointment 0.5 ml to each incision.

Bandage Enhancement Experiment

Controls – Hydrogel pads without materials or ointment added to the pad will be used as the control.

Constants – The amount of colloidal silver injected into the hydrogel will be kept at 1.0 ml; the temperature of the conditions will be 25oC; the size of the hydrogel pads will be kept at 7 cm x 5 cm.

(9) What was your experimental process? Include each of the steps in your experiment.



** Please open the PDF file titled “Experiments” to see photos of the team working on all three experiments, plus explanations, procedures, and data analysis for every experiment.

In testing bacterial growth, the team was resourceful in identifying affordable ingredients that were antimicrobial. Colloidal silver, probiotics, and xylitol are common substances that can be purchased over-the-counter by any consumer. Each of these materials, plus the unique and novel “colloidal silver + xylitol” composite that will be created by the team, will be tested in vitro experimentation using bacteria-inoculated agar in Petri dishes. Circular disks of filter paper will be saturated with each solution and then placed in Petri dishes and incubated at 37oC for 16 hours. The team will measure the diameter of the disk outward to the closest bacterial colony. This area free from bacteria surrounding the disk is called the “zone of inhibition.” The greater the zone, the more effective the material was in fighting biofilms, and the closer the team would be to identifying an alternative solution for chronic wounds.

** This process will be repeated three times for reliability. Specific procedures are listed below to allow anyone to replicate this experiment and duplicate the results of these tests for verification.

In Vitro Experiment -

To prepare Petri dishes for inoculation, use this procedure:

1) Weighed out the following into a 1 L flask:6 grams agar250 ml of distilled water 2) Heat to boiling for one minute in a pan placed on a hotplate3) Stir to mix and dissolve the agar.4) Pour agar until it just covers the bottom of Petri dishes.5) Place lid on Petri dishes, invert, and allow to cool at room temperature.

To test the antibacterial disks against bacteria, use this procedure:

1) Cut circular disks out of filter paper soaked with the solution used – silver, xylitol, probiotic, or silver + xylitol – a composite material.2) Measure to ensure each disk is 5 mm in diameter.3) Label each Petri dish with the solution being tested – silver, xylitol, probiotic, or silver + xylitol – a composite material.4) As directed by Dr. Rumbaugh of the Texas Tech Medical School, gently touch the agar in the dish to inoculate with bacteria common to skin. Give one short cough on the agar to inoculate with airborne bacteria.5) Divide each Petri dish into quadrants and place four disks, each soaked with a solution, into the separate sections of the plate.6) Place Petri dishes into an incubator, inverted to prevent condensation from forming on the agar.7) Set incubator to body temperature, 37o Celsius.8) Measure the zones of inhibition (mm) surrounding each disk after 16 hours.9) Analyze the most effective material by looking at the data for the disk with the greatest average zone of inhibition.10) Repeat this procedure on two additional sets of experiments.11) Use the material identified as most effective in the following in vivo experiment.

In Vivo Experimentation

** To test whether or not colloidal silver would be successful when applied to an incision on an earthworm, the team used the novel idea of combining pyranine, a chemical commonly found in highlighter markers and which is non-toxic, with the colloidal silver so that the incisions on the earthworms could be seen more easily. Without the use of pyranine under the black light, it was difficult to locate and measure the incisions.

1) Obtain 20 earthworms, species Lumbricus terrestris. 2) Prepare four containers with soil for housing earthworms, labeled A, B, C, D.3) Place three uncut worms into container A, as a control. Label.4) Cut an incision 1.0 cm in length below the clitellum band on each of nine earthworms. 5) In a 40 ml beaker, add the felt of a yellow highlighter marker to extract the chemical pyranine. (non- toxic, glows under black light)6) Add 0.5 ml of colloidal silver to an equal amount of pyranine.7) With the exception of our control experiment, each worm’s incision was treated with colloidal silver using a sterile swab.8) Once the solution is applied, each worm is put into its own container and set aside overnight.9) Under black lights, remove worms from soil and locate the incision as it glows in the dark.10) Each day the worms’ incisions will be measured in millimeters.11) Repeat the in vivo experiment two times for reliability. 12) Data will be analyzed to determine if the colloidal silver increased the rate in which the incisions healed.

Bandage Enhancement

When testing the effect of colloidal silver when applied to a hydrogel bandage, use this procedure.

Hydrogel Test:1) A syringe was filled with 0.5 ml of colloidal silver2) The hydrogel was peeled back and the silver was injected into the side of the dressing. Repeat on two other pads.3) Measure the amount of material that enters the hydrogel in milliliters.4) Observe hydrogel pads daily for leakage.5) Collect qualitative data including moisture level and amount of silver seen inside the transparent hydrogel.6) Collect quantitative data – amount of colloidal silver leaking out of hydrogel.7) Repeat experiment two additional times for reliable results.8) Analyze qualitative and quantitative data.

Data Collection and Analysis

(10) Describe the data you collected and observed in your testing (use of data tables, charts, and/or graph is encouraged).

** Please see the three uploaded ExCel spreadsheets titled “In Vitro Experiment Data,” “In Vivo Experiment Data,” and “Bandage Enhancement Data,” that contain data tables, charts, and bar graphs (including explanations) of the results of the experiments.



** Please also open the PDF file titled “Experiments” to see photos of the team working on all three experiments, plus explanations, procedures, and data analysis for every experiment.

In Vitro Experiment – Colloidal silver consistently produced the largest zones of inhibition surrounding the treated disks when compared with the other experimental groups. In all three tests and in every trial within each test, the disk treated with silver alone was effective in fighting biofilms and bacteria.

In Vivo Experiment – Since silver was the standout during in vitro testing for all three experiments, it was used during in vivo testing on earthworms. Again, it was very successful in promoting cell regrowth and closing the incision wound of the worm in a short amount of time.

Bandage Enhancement – With the success of silver fighting bacteria off in vitro and promoting cell regrowth in vivo experimentation, it was used as an added material to hydrogel pads. The injections were effective in keeping the medication as part of the bandage which would continually release the colloidal silver onto wounds.

(11) Analyze the data you collected and observed in your testing. Does your data support or refute your hypothesis? Do not answer with a yes or no. Explain your answer using one of the following prompts: 'Our data supports/refutes the hypothesis because...'

In Vitro Testing:

The data refutes the hypothesis that stated, the composite material of silver plus xylitol we created would show greater ability to fight bacterial colonies. In the experiment, colloidal silver by itself was most effective in preventing bacterial colonies from overtaking the test materials. Silver had the largest zone of inhibition surrounding the test disks in the Petri dishes, which means the biofilms detected silver and could not grow in its presence.

In Vivo Testing -

The data from this experiment supports the hypothesis stating that silver would be one of the best candidates for encouraging cell regrowth and closing wounds. The colloidal silver prevented the bacteria from making a biofilm over the incisions on the earthworms and healing occurred under the experimental conditions.

Bandage Enhancement -

The data collected during the bioengineering of a bandage supported the hypothesis. Hydrogel pads were able to be embedded with colloidal silver and the material remained in the bandage throughout testing. With a bandage that maintains the ointment inside the pad, gradual release will allow bacteria to be reduced throughout the process, and less changes of bandages is needed with moist hydrogels.

(12) Explain any sources of error and how these could have affected your results.

** Please see the uploaded data table titled, "Experimental Errors"

Every experiment has sources of error and although good scientists try to keep these to a minimum, they do affect results.

During the bacterial growth experiments, the saturated filter made punching out disks a disadvantage. Cutting out the disks from the filter paper prior to its saturation proved to be much easier. The size and shape of the disks were more precise and constant during experimentation. Before applying the saturated filters to the agar, allow them to dry out. Changing the procedure resulted in more accurate measurements in millimeters during the zone of inhibition data collection.

A systemic error we experienced was setting the Petri dishes in the incubator face up during the trial run before actual experimentation began. Because the Petri dish was face up, condensation was collecting at the top of the dish, not allowing us to observe the inside clearly. After consulting with medical students at Texas Tech we learned for a more accurate outcome, to put the Petri dishes in the incubator face down. Once we began the actual experiment, it was easier to evaluate zones of inhibition through a clear lid.

A systemic error made was using an incubator that is not new and did not always keep the environment at 37oC – it fluctuated from 36o to 38o Celsius. After redoing the experimentation, the results proved to be repeatable and valid.

Drawing Conclusions

(13) Interpret and evaluate your results and write a conclusion statement that includes the following: Describe what you would do if you wanted to retest or further test your hypothesis. Evaluate the usefulness of the data your team collected. What changes would you make to your hypothesis and/or experimental design in the future, if any?

The result of team Silver Bullet is exciting. The colloidal silver used during experimentation was by far the most successful, common, affordable ingredient in fighting bacteria and was a key component in destroying biofilms. Biofilms plague the planet - causing slow healing, blood clots, infections leading to amputations, and heart disease. Through this project, the true colors of biofilms were seen and defeated in preliminary testing.

It is important to continue searching for treatments to conquer chronic wounds because of the incredible number of people who suffer.

To further test the hypothesis, colloidal silver should be combined with other materials and composites tested. At the nano level, silver could be used internally and the fight could result from the inside out. Gold nanoparticles have shown antimicrobial promise, so if combined with silver, in vitro and in vivo testing could result in an even more effective treatment.

A new hypothesis would be tested as well. Certain essential oils (frankincense, cedar) are also showing evidence of killing bacteria resistant to over-prescribed antibiotics. Using these in testing would allow for collaborations with many organizations today that are popularizing the use of oils and diffusers.

Working with Dr. Kendra Rumbaugh, a biofilms expert at Texas Tech and globally recognized for her work with biofilms and mice, the tests performed on invertebrate earthworms could be tested under controlled conditions in her laboratory using vertebrates. This next step of testing would be important in bringing a solution to reality. In fact, the data collected by our team is useful only because of our connections with doctors working in clinics and medical schools whose focus is on chronic wounds. Our experiment was not performed on an island – the information we learned was requested by these leaders in the field of wound care because they recognize that alternative treatments are essential in a world steeped in antibiotic-resistance.

The experiments conducted over and over led to better techniques in testing and more reliable results for society. Changes in our procedures for the future include experimenting with different concentrations of colloidal silver to find the most ideal percentage while balancing affordability.



Uploaded Files:

• [ View ] Contact List (By: anthrobones, 02/25/2016, .pdf)

Major supporters of the team this year are included in this list. They helped by allowing us to visit their laboratories and they gave us

advice on how to best test for the growth of bacteria in vitro.

• [ View ] Collaboration with Community Experts (By: anthrobones, 02/26/2016, .pdf)

Work with many regional experts in chronic wounds and biofilms is shown in this pdf file. Included are notes from interviews,

brainstorming pages from the team, photos of interviews and lab training, and exciting collaborations with experts in the community and

beyond.

• [ View ] Bibliography (By: anthrobones, 02/27/2016, .pdf)

Bibliography includes websites, professional journals, personal interviewees, newspaper & magazine articles, TED talks, webinars, and

relevant organizations and university resources.

• [ View ] In Vitro Experiments Data (By: anthrobones, 02/29/2016, .xlsx)

Data Tables, charts, graphs, and explanations of the analysis are found in this ExCel spreadsheet. They are also included in the PDF

upload titled "Experiments."

• [ View ] Data Table - Experimental Errors (By: anthrobones, 02/29/2016, .pdf)

This data table accompanies the answers to the question about errors that may have affected the experiments.

• [ View ] In Vivo Experiments Data (By: anthrobones, 02/29/2016, .xlsx)

In Vivo experiments are those conducted on living organisms. INVERTEBRATES ONLY WERE USED DURING THIS PROJECT,

THEREFORE NO VERTEBRATE FORM IS REQUIRED. This ExCel spreadsheet shows the data collected and analyzed, has an

explanation included, and contains data tables, charts and graphs for all three trials of all three experiments conducted using

earthworms and silver.

• [ View ] Bandage Experiment Data (By: anthrobones, 02/29/2016, .xlsx)

The ability to load colloidal silver into hydrogel using a sterile needle and syringe was an important test to see if there could be a

solution in the form of an enhanced bandage. This ExCel spreadsheet shows a table and explanation of the 5 trials conducted. It can

also be found in the Experiments PDF Upload containing photos and data from every experiment.

• [ View ] Experiments (By: anthrobones, 02/29/2016, .pdf)

A MUST-SEE Please open this PDF file converted from PowerPoint that includes many photos of the team working on all three

experiments. Steps are included, data tables, graphs, charts, explanations of data, and the story of this project can be seen in this

document. A MUST-SEE

• [ View ] Bacteria Experiment Lab Report (By: anthrobones, 02/29/2016, .pptx)

Lab report with details about the bacteria experiment.

Community Benefit(1) How could your experiments and data help solve your problem and benefit your community? Describe next steps for further research/experimentation and how you have or how you could implement your solution in the future.

The data and experiments could solve the problem of chronic wounds because a material was identified that is already FDA approved for topical use and is an affordable solution that kills bacteria. An ointment and hydrogel bandage was bioengineered for treating chronic wounds in not just this community, but in the nation. In our community, the oil field and agriculture are responsible for the predominant economy, yet they are the two most dangerous fields of work in the West. Numerous injuries are reported every year due to these occupations. Combined with diabetes which is a leading cause of death in our county, chronic wounds are far too common. Our colloidal silver hydrogel-bandage will prevent dangerous bacterial infections in chronic wounds if further experimentation leads this data to a university laboratory and medical school. Implementing the solution in the future will only result if the project continues past the competition deadline. Silver Bullet is more than a short term project; we are interested in this solution for the long run. Our community has benefited from our data and experiments because of the public awareness campaigns of which we are a part. People with diabetes have been targeted as those with great interest in this solution. After consulting their doctor, the ointment we recommend from data analysis would be affordable in this county and is available locally.

The following is a list of ways our team has implemented the solution and is making a difference in the community and changing the results of chronic wounds:

• IRB approval to test colloidal silver on vertebrates through Texas Tech• Preliminary results already reported to Dr. Rumbaugh in the medical school• Presentation to the Cochran County Senior Citizens Center• Presentation to the Community Health Fair• Develop a commercial name for the ointment of colloidal silver• Manufacture colloidal silver-embedded bandage of hydrogel• Reports to all doctors who advised this team• April poster presentation given alongside graduate students at Texas Tech• Website is continuing to expand the information as pages are added • Media from the regional newspapers are reporting our story of success• Certain geometric shapes from nature (biomimicry) may prevent bacteria from spreading deeper into wounds, therefore bandage shape is a new development

Our team knows we chose a topic this year that is a great challenge. There is not one answer. There is no easy answer. There are many searching for the solution. That doesn’t mean people like us shouldn’t join the fight.

To summarize this project:

Chronic wounds have become a significant health issue and a $25 billion business in the United States. With an aging population and a steep rise in diabetes and obesity, heartbreaking wound complications can result in blood clots, heart disease, and amputation. The goal of cost effective ways to heal wounds in a world with antibiotic-resistant bacteria was the team challenge. Research and interviews led to experiments using xylitol, probiotics, and colloidal silver as possible ways to attack biofilms in chronic wounds.



The team tested the effectiveness of these materials individually and created composite materials. In vitro tests used Petri dishes of nutrient agar inoculated with airborne bacteria and colonies from direct contact. The results identified colloidal silver as the material with the largest zone of inhibition and greatest ability to fight bacteria. On treated discs, bacteria could not form at all, and in tests with established bacterial colonies, silver fought back. It was the most cost effective healing substance at approximately one dollar per application, as opposed to much higher costs for gold nanoparticle ointments or oxygen chamber treatments. During in vivo invertebrate testing, silver again showed great promise by speeding cell growth and closing wounds on earthworms. When silver was injected into hydrogel, a water-based pad, its potential was fully realized as a bandage with benefits of moisture, comfort, healing properties of colloidal silver, and affordability. Working with university research scientists, this STEM solution is designed to benefit the 6.5 billion people suffering from chronic wounds.

The battle with chronic wounds continues, but with the information learned this year, we are one step closer to conquering the ugliness of wounds that refuse to heal. “We don’t grow when things are easy. We grow when we face challenges.” – Joyce Meyer

Uploaded Files:

• [ View ] Website URL and QR Code (By: anthrobones, 02/28/2016, .pdf)

The Silver Bullet website spreads awareness of chronic wounds and alternative treatments becoming available. The website's URL and

QR code are included in this document.

• [ View ] Community Health Fair (By: anthrobones, 02/28/2016, .pdf)

A flyer was distributed to raise awareness of the health fair and information the team would present on "Middle School STEM

Involvement and Chronic Wounds." There are alternative treatments for chronic wounds of which many people are unaware. Silver

Bullet aims to change that.

• [ View ] Senior Citizens Presentation (By: anthrobones, 02/28/2016, .pdf)

A flyer advertising the presentation from the Silver Bullet team to the senior citizens meeting of Cochran County is included in the

uploads. Some of the most vulnerable people for chronic wounds live in our county and education here is important for them.

Mission Verification(1) Does your Mission Folder project involve vertebrate testing, defined as animals with backbones and spinal columns (which include humans)? If yes, team must complete and attach an IRB approval form.

No

(2) Did your team use a survey for any part of your project? If yes, team must complete and attach a survey approval form.

No

(3) You will need to include an abstract of 250 words or less. As part of the abstract you will need to describe your project and explain how you used STEM (Science, Technology, Engineering and Mathematics) to improve your community

Chronic wounds have become a significant health issue and a $25 billion business in the United States. With an aging population and a steep rise in diabetes and obesity, heartbreaking wound complications can result in blood clots, heart disease, and amputation. The goal of cost effective ways to heal wounds in a world with antibiotic-resistant bacteria was the team challenge. Research and interviews led to experiments using xylitol, probiotics, and colloidal silver as possible ways to attack biofilms in chronic wounds.The team tested the effectiveness of these materials individually and created composite materials. In vitro tests used Petri dishes of nutrient agar inoculated with airborne bacteria and colonies from direct contact. The results identified colloidal silver as the material with the largest zone of inhibition and greatest ability to fight bacteria. On treated discs, bacteria could not form at all, and in tests with established bacterial colonies, silver fought back. It was the most cost effective healing substance at approximately one dollar per application, as opposed to much higher costs for gold nanoparticle ointments or oxygen chamber treatments. During in vivo invertebrate testing, silver again showed great promise by speeding cell growth and closing wounds on earthworms. When silver was injected into hydrogel, a water-based pad, its potential was fully realized as a bandage with benefits of moisture, comfort, healing properties of colloidal silver, and affordability. Working with university research scientists, this STEM solution is designed to benefit the 6.5 billion people suffering from chronic wounds.

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Silver Bullet Join the fight against biofilms and prevent infection in chronic wounds

Blade Kaden Kaylah Elizabeth

EnthusiasmForTheProjectCoordinatedthe

BrainstormingSessions

RecordsNotesduringInterviewswith

CommunityExperts

CommunicationSpecialist

StrongLeadershipSkills

WorksTogetherwellwithAnyoneGoodListener

InterviewingSpecialistWritesThank-younotes

ProblemSolver/TroubleShooter

Askstoughquestions

EnjoysExperimentsGoodLabManager

Graphs,Charts,DataGenerator

DataCollection&Measurements

ThoroughResearcher StrongLabScientist TimeManagementSpecialist

PublicSpeakerduringPresentations

IdeaGeneratorStudiesPaleontology

StrongWriterCoordinatedOutreach

withDr.Lehman

StrongWritingSkillsOrganizerforMeetings

SeestheBigPictureEasily

GrandIdeasanddreams

InnovatorandCreative

AdvancedMathematician

InteractswellwithMedicalScientistsHighlyVerbal

StrongAcademics

PreparedSupportMaterials

Artist

ThoroughduringResearch&Summaries

DrivenandDeterminedPosterPresentations

OrganizedOnTime

HighVocabularyinVerbalandWrittenCommunication

Takesnotesduringinterviews

Detail-orientedAdvanceCalculator

Skills

AnalyzesDataWellDrawsConclusions

IndependentWorkerHighlyDependable

EncouragertotheTeamandanOutgoingTeam

Member

PhysicallyOutgoingFlexible&Compromises

DataCollectionCreatedTables&

QualitativeDataSheets

AcademicallyAdvancedMemoryforDetails

GoodListenerAppreciativeofOthers

Ideas

PreparedPowerPointsandPresentationsThoughtofourTeamName

SociallyOutgoing

WorkswellwithOtherPeopleofallAges

Logical&aVoiceofReasonfortheTeamKeepsothersonTrack

August2015May2015 Sept2015 Oct2015April2015 Nov2015

Research&topicchosen:Waystoeffectivelybreakdownbacteriathatwouldbecosteffective.

EcybermissionRegistration:TeamwascompletedwithmembersElizabeth,Kaylah,Kaden,andBlade.

BeganExperimentation:Chosecosteffectivesubstanceswithhealingproperties.

TeamandActionPlanDevelopment TeamMeetings– ActionPlan

DevelopedChoseascheduleforcompletionofMissionFolder&Experiments

ConsultedwithJosieMusicoSheinterviewedustospreadthenewsofwoundhealing.

Silver BulletTimelinefor eCYBERMISSION Project

April2015~May2016

Feb2016 March2016 April2016Jan2016 May2016

ContactedOtherExpertsConductedtestswithmedicalresearchers,Dr.KendraRumbaugh,RandallWolcott,intheBiopolymerInstitute,MedicalBiofilmResearchInstituteandTexasTechHealthandSciencesCenter.

PosterPresentationPrepareposterforTexasTechUniversityGraduateSchool.

Testedthechosensubstancestobreakdownthebacteria

FuturePlansfortheProjectistodevelopahydrogelbandagethatwilldeliverthecolloidalsilverdirectlyandeffectivelytothewound

Silver BulletTimelineContinued ...

Set up interviews with professional doctors. Continued doing research for the project Silver Bullet.

Start a public awareness campaign about wound care.

Contact List for Silver Bullet

Majorsupporters,subjectmatterexperts,andcollaborationsduringthisprojectarelistedbelowwithcontactinformationforjudges’convenience.Onlythosepeoplewith

whomtheteamhadcontinuouscontactareincludedonthislist.GregoryRogers,MDCovenantHospitalLevellandFamilyandEmergencyMedicine1900CollegeAvenueLevelland,Texas79336806-894-4963

KendraRumbaugh,Ph.D.AssociateProfessorDepartmentofSurgery,MS8312TexasTechUniversityHealthSciencesCenter36014thStreetLubbock,[email protected],MDSouthwestRegionalWoundCenter2002OxfordAvenueLubbock,TX79410(806)793-8869MichaelLehman,MDLehmanDermatology371521stStreetLubbock,Texas79410806-795-0617

Manythankstotheindividualswhoallowedustovisitwiththemabouttheissueofchronicwounds.Theiridentityisbeingkept

hiddenforprivacy.

C H R O N I C W O U N D S / B I O F I L M SC O M M U N I T Y E X P E R T S

S I LV E R B U L L E TE C Y B E R M I S S I O N 2 0 1 6

A collection of interviews and collaborations between the team and regional experts

Questions Answers Teamwork Photos Lab Experiences Advice

INTERVIEWS & COLLABORATIONS• Dr. Michael Lehman, MD Lehman Dermatology, Lubbock, Texas – Wound & Skin Disease Specialist

• Dr. Angel Cueva, Texas Tech Medical School, Lubbock, Texas – Laboratory research, advanced technology

• Dr. Hannah Zhao, Texas Tech University Health Sciences Center, Department of Surgery, Lubbock, Texas, Microbiologist

• Dr. Greg Rogers, MD – Lab and medical techniques, General Physician, Covenant Hospital, Levelland, Texas

• Michael Briggs, Neuropathy patient with chronic wounds, Lubbock, Texas

• Dr. Randy Wolcott, MD – Southwest Regional Wound Care Center, chronic wound care specialist, Lubbock, Texas

• Dr. Kendra Rumbaugh, PhD, Texas Tech University Health Sciences Center, Microbiologist, Department of Surgery, Texas Tech Medical School, Chronic Wounds Laboratory, Lubbock, Texas

INTERVIEW NOTES TAKEN BY THE TEAM

QUESTIONS AND BRAINSTORMINGBY THE TEAM

COLLABORATION PHOTOS

DR. ROGERS

Dr. Rogers had expertise in dealing with chronic wounds because he has many

patients in rural clinics with diabetic

complications.

Dr. Rogers helped us understand the

difference between a wound, a serious wound,

and a chronic wound that can be fatal. He answered all of the

questions we prepared in advance.

Silver Bullet team invited Dr. Rogers to the science lab to see the progress of the in vitro

experiments and ask advice on in vivo experimentation.


DR. LEHMAN

Our team asked Dr. Lehman for advice

about our methods of engineering a bandage

to fight biofilms in chronic wounds.

Dr. Lehman demonstrated the

proper way to apply a moist bandage to

prevent infection. We wanted to be certain we

followed proper protocols

Kaylah and Dr. Lehman in the dermatology clinic after discussing wound care.


DR. ZHAO

Dr. Zhao described types of beneficial and

dangerous bacteria – and how the LOCATION of that bacteria makes the

difference.Notetaking – a real

world skill


DR. WOLCOTT

Learning how to disrupt a matrix of biofilms that allow bacteria to communicate. If silver and xylitol can stop

the communication, wounds could heal.

The Silver Bullet team with Dr. Wolcott in the clinic learning about cutting edge research in

the field of chronic wounds.


DR. WOLCOTTDr. Wolcott’s poster on

Disrupting Biofilm Matrices – the very issue

we were studying.

Demonstrating the protocols for testing materials and their

abilities to fight infection in vitro experiments.

Technology such as chambers of oxygen and ozone are also being used to treat chronic

wounds that do not respond to medications.


DR. CUEVA

Dr. Cueva taught us to load and deliver liquids

through a pipette

Practicing the use of a pipette to make

concentration gradients

Technology such as chambers of oxygen and ozone are also being used to treat chronic

wounds that do not respond to medications.

COLL ABORATION PHOTOS

DR. CUEVADR. RICHARDS

Dr. Wolcott’s poster on Disrupting Biofilm

Matrices – the very issue we were studying.

Always ask a university if you need the use of better and more

advanced equipment than

what your school can provide! They

say YES!

Practice makes perfect


DR. RUMBAUGH’S LAB

Ecybermission led us to a fantastic role model

We were so excited to be able to work with Dr. Kendra

Rumbaughwho is a global

leader in the field of

biofilms and

chronic wounds!

Bibliography

Anthes, E. 7 Next-Gen Bandages That Help Heal Wounds. Retrieved August 20, 2015. Auddy, Runa. "New Guar Biopolymer Silver Nanocomposites for Wound Healing

Applications." Http://www.hindawi.com/journals/bmri/2013/912458. BioMed Research International, 18 Aug. 2013. Web. 4 Oct. 2015.

Brickey, Henri. "Trials to Showcase Power of Phages in Fighting Infection." Lubbock Avalanche-Journal 4 Jan. 2007. Print. Cardinal, Matthew, David E. Eisenbud, and David G. Armstrong. "Wound Shape Geometry

Measurements Correlate to Eventual Wound Healing." Wound Repair and Regeneration 17.2 (2009): 173-78.

Cevher, E. (2011, November 16). Biopolymers as Wound Healing Materials: Challenges and New Strategies. Retrieved November 16, 2015. Cueva, Angel, MD. “The Techniques of using Pipettes for Quantifying Research.” Personal

interview conducted by Blade Henry, Kaden Moses, Kaylah Nunn, and Elizabeth Casarez. 12 Feb 2016.

Dissanaike, Sharmila, MD. "TTUHSC Surgery Burn Center of Research Excellence (BCoRE)." TTUHSC. 13 Dec. 2015. Donlan, Rodney M. "Biofilms: Microbial Life on Surfaces." www.ncbi.nih.gov. Rodney M. Donlan, 13 Dec. 2015. Web. 25 Feb. 2016. Engelking, Carl. "Nanotech Lotion Speeds Diabetic Wound Healing." Discover Magazine. Kalmbach Publishing, April 2016 Issue: Subscribers Sneak Peek. Web. 10 Feb. 2016. Fleming, Derek, MD. “The Role of a Medical Technologist.” Personal Interview conducted by Kaden Moses and Blade Henry. 12 Feb 2016. Harada, Cesar. "How I Teach Kids to Love Science." TEDX. N.p., n.d. Web. 4 Sept. 2015. Lehman, Michael, MD. “Antibiotic Resistance and Treating Biofilms for Success in Healing Chronic Wounds.” Personal Interview by Kaylah Nunn. 17 Jan 2015. Martin, Normin. "Yang Hu Takes New Post with Tech’s Department of Plant and Soil Science." www.depts.ttu.edu. CASNR News Center, 13 Dec. 2015. Web. 25 Feb. 2016. Nightingale, Sarah. "Show 'Health Care Heroes' Features Lubbock Doctors Known for Treating

Hard-to-heal Wounds." Lubbock Avalanche-Journal 8 Sept. 2010. Print. Paddock, Catharine, PhD. "MRSA / Drug Resistance Infectious Diseases / Bacteria / Viruses

Anti-biofilm Molecule May Help Fight Superbugs." Medical News Today. MNT, 23 May 2014. Web. 8 Sept. 2015.

Pastore, Rose. "How A Simple New Invention Seals A Gunshot Wound In 15 Seconds." Popular

Science. Bonnier Corporation Company, 03 Feb. 2014. Web. 12 July 2015. Richards, Summer, MD. "Biofilms and Their Role in Infection." Personal interview conducted by Kaylah Deavours Nunn. 4 Feb. 2016. Rogers, Gregory, MD. “Prevalence of Chronic Wounds in Emergency Rooms and Clinics of

Rural West Texas.” Personal Interview conducted by Kaylah Nunn, Blade Henry, Elizabeth Casarez, Kaden Moses. 10 Oct 2015.

Rumbaugh, Kendra. "Department of Surgery - The Rumbaugh Lab." www.ttuhsc.edu. Kendra Rumbaugh, 13 Dec. 2015. Web. 02 Oct 2015. Sezer, Ali. "Biopolymers as Wound Healing Materials: Challenges and New Strategies." www.intechopen.com. InTech, 16 Nov. 2011. Web. 4 Oct. 2015. Venkatrajah, B. "Biopolymer and Bletilla Striata Herbal Extract Coated Cotton Gauze

Preparation for Wound Healing." http://scialert.net PDF, 24 Apr. 2013. Web. 4 Oct. 2015.

Wolcott, Randy, MD. “Treatment Alternatives for Chronic Wounds: Fighting Biofilms.” Interview by Kaylah Nunn, Elizabeth Casarez, Kaden Moses, Blade Henry. 21 Jan 2015. Zhao, Hannah, MD. “Preparing Petri Dishes for Bacterial Growth and Analysis.” Personal Interview conducted by Kaylah Nunn and Elizabeth Casarez. 12 Feb 2016. Zhao, X. PhD. Stretchable Hydrogel Electronics: Water-based “Band-Aid”

senses temperature, lights up, and delivers medicine to the skin.” MIT News. Retrieved December 7, 2015.

Sample 1 Sample 2 Sample 3

Probiotics 9 8 6

Xylitol 4 12 5

Silver 15 15 5

Composite - Silver + Xylitol 6 12 4

Control 5 3 2


Probiotics 6 8 5

Xylitol 4 11 6

Silver 20 25 15


Control 5 3 2

Testing the Effectiveness of Materials in Preventing Bacterial Growth In Vitro

MaterialSize of Zone of Inhibition Surrounding the Disk (mm)

Trial 1


Trial 2

Silver was the most effective ingredient tested in vitro for stopping bacterial growth. In Trial 1, Trial 2, and Trial 3 consistent results were obtained in sample after sample tested. Our hypothesis was refuted because we expected the composite of silver + xylitol to "outperform" silver treatments alone. Based on the results of repeated testing, silver will now be tested in vivo to learn if similar success in fighting bacteria and promoting cell regrowth/healing will occur on a living organism. Testing will NOT involve vertebrates will be used for experimentation.


Probiotics 7 7 10

Xylitol 2 7 9

Silver 24 23 19


Control 2 3 8

Trial 3


Sample 4 Mean

3 6.5

3 6

10 11.25

5 6.8

8 4.5

Sample 4 Mean

5 6

7 7

20 20

13 8.8

8 4.5

Testing the Effectiveness of Materials in Preventing Bacterial Growth In Vitro

Size of Zone of Inhibition Surrounding the Disk (mm)

Trial 1


Trial 2

in vitro for stopping bacterial growth. In Trial 1, Trial 2, and Trial 3 consistent results were obtained in sample after sample tested. Our hypothesis was refuted because we expected the composite of silver + xylitol to "outperform" silver treatments alone. Based on the results of repeated testing, silver will now be tested in vivo to learn if similar success in fighting bacteria and promoting cell regrowth/healing will occur on a living organism. Testing will NOT involve vertebrates - invertebrates only

0 2 4 6 8 10 12

Probiotics

Xylitol

Silver

Composite - Silver + Xylitol

Control

Effective Materials for Preventing Bacterial Growth: Trial 1

Zone of Inhibition (millimeters)

Probiotics

Xylitol

Silver


Control


Sample 4 Mean

2 6.5

8 6.5

22 17.6

12 7

3 4

Trial 3


0 5 10 15 20 25

Probiotics


0 2 4 6 8 10 12 14 16 18 20

Probiotics

Xylitol

Silver


Control



SourcesofExperimentalErrors

SourceofError ExperimentAffected PossibleErrorRecorded

ChangesthatCouldImproveResultsin

theFuture

Controlofincisionstotheearthworms InVivoTesting

Incisionsmadeontheearthwormswereapproximatelythesamelengthbutnot

exact

Useofadissectingmicroscopeforbetterviewing;remeasuring

incisionmultipletimesasthe

earthwormstretchedandcontracted

Ointmentappliedtoearthwormswith

incisionsInVivoTesting

Noointmentwasappliedtothecontrol

earthworm

Useanointmentwithoutanactiveingredientsuchas

Vaselinenexttimetocomparewiththe

ointmentwithtestedingredients

Incubatortemperature InVitrotesting

Temperaturewassetfor37oCorbodytemperaturebut

fluctuatedbetween36oand38oC

Conductinvitroexperimentsinauniversitylabwithnewerincubators

Earthworm 1 Earthworm 2 Earthworm 3 Earthworm 1 Earthworm 2Control No Silver 1 1 0.8 0.7 0.8

Colloidal Silver 0.7 0.7 0.6 0.3 0.4

Earthworm 1 Earthworm 2 Earthworm 3 Earthworm 1 Earthworm 2Control No Silver 0.9 0.8 1 0.7 0.5


Earthworm 1 Earthworm 2 Earthworm 3 Earthworm 1 Earthworm 2Control No Silver 0.9 1 0.7 0.8 0.8


Condition Applied Day 3 Day 4

In Vivo Experimentation - Test 1 - Measurement of I Day 3 Day 4Condition Applied

In Vivo Experimentation - Test 2 - Measurement of I Condition Applied Day 3 Day 4

In Vivo Experimentation - Test 3 - Measurement of

0

0.5

1

1.5

1 2 3

Mean Results Test 1: Control No Silver

0

0.5

1

1

Mean

0

0.2

0.4

0.6

0.8

1

1 2 3


0

0.2

0.4

0.6

0.8

0

0.2

0.4

0.6

0.8

1 2 3

Mean Result Test 3: Control No Silver

0

0.2

0.4

0.6

0.8

Earthworm 3 Earthworm 1 Earthworm 2 Earthworm 30.8 0.5 0.5 0.40.5 0 0.1 0.2

Earthworm 3 Earthworm 1 Earthworm 2 Earthworm 30.7 0.5 0.4 0.50.2 0 0 0

Earthworm 3 Earthworm 1 Earthworm 2 Earthworm 30.7 0.6 0.4 0.50.2 0.1 0.2 0

Day 5

ncision (Initial = 1.0 cm) Day 5

ncision (Initial = 1.0 cm) Day 5

Incision Initial = 1.0 cm

1 2 3

n Results Test 1: Colloidal Silver

1 2 3

Mean Results Test 2: Colloidal Silver

1 2 3

Mean Result Test 3: Colloidal Silver

During tests conducted in vivo using the earthworm Lubricus terrestris for testing, a 1 cm incision was made into the skin of the worm just below the clitellum band. Silver was mixed with pyranine, a non-toxic chemical that glows brilliantly under UV black light. Under a black light, it was easy to see the site of the incision and make measurements, whereas it was extremely difficult before this treatment. Then, the silver was applied to the wound. Earthworms were returned to their containers and measurements were made in cm on Day 3, 4, and 5. The worms treated with silver had greater cell growth and wound healing faster than those with no silver applied. These results were consistent during testing and show that colloidal silver does speed cell regrowth and healing on living organisms.

Trial Amount of Silver in Syringe (ml)1 0.52 0.53 0.54 0.55 0.5

In every test except Test 3, 100% of the silver that wa transferred into the hydrogel pad. The silver remaine testing. This hydrogel enhanced bandage with colloid airborne bacteria, and in contact with colloidal silver

Percent of Silver Successfully Transferred to Hydrogel (ml)100%100%85%

100%100%

as contained in the syringe was able to be successfully ed in the hydrogel as part of the bandage throughout

dal silver would keep a wound moist, protected from during the time the bandage remains on the wound.

In Vitro Experiments

Photos * Data * Tables * Graphs

Silver Bullet Ecybermission Team 2016

Preparing Petri dishes with agar and placing disks to fight bacteria

Labelling Petri dishes and inoculating with probiotics, colloidal silver, xylitol, and silver/xylitol composite.

Placing Petri dishes into the incubator and setting the thermometer to control the experiment at body temperature or 37o C

Ready to measure the zones of inhibition using millimeters.

Zones of Inhibition were measured (mm) from the center of the paper disks outward to the nearest bacterial colony.

TestingtheEffectivenessofMaterials inPreventingBacterialGrowthInVitroTrial1

Material Sample1 Sample2 Sample3 Sample4 MeanSizeofZoneofInhibitionSurroundingtheDisk(mm)

Probiotics 9 8 6 3 6.5Xylitol 4 12 5 3 6Silver 15 15 5 10 11.25

Composite- Silver+Xylitol 6 12 4 5 6.8Control 5 3 2 8 4.5

0 2 4 6 8 10 12

Probiotics

Xylitol

Silver


Control


Zone of Inhibition (mm)

Trial2


Probiotics 6 8 5 5 6Xylitol 4 11 6 7 7Silver 20 25 15 20 20

Composite- Silver+Xylitol 3 12 7 13 8.8Control 5 3 2 8 4.5

0 5 10 15 20 25

Probiotics

Xylitol

Silver


Control



Trial3


Probiotics 7 7 10 2 6.5Xylitol 2 7 9 8 6.5Silver 24 23 19 22 17.6

Composite- Silver+Xylitol 1 9 6 12 7Control 2 3 8 3 4

0 2 4 6 8 10 12 14 16 18 20

Probiotics

Xylitol

Silver


Control



Colloidal silver was the most effective in stopping bacterial growth. Since it was successful during in vitro testing, it would be used during in vivo testing.

Image from www.rochway.com.au

Silver was the most effective ingredient tested in vitro for stopping bacterial growth. In Trial 1, Trial 2, and Trial 3 consistent results were obtained in sample after sample tested. Our hypothesis was refuted because we expected the composite of silver + xylitol to "outperform" silver treatments alone. Based on the results of repeated testing, silver will now be tested in vivo to learn if similar success in fighting bacteria and promoting cell regrowth/healing will occur on a living organism. Testing will NOT involve vertebrates -invertebrates only will be used for experimentation.

In Vivo Experiments

Photos * Data * Tables * Graphs

Silver Bullet Ecybermission Team 2016

Working together to make a 1 cm incision in the earthworm

Sunday afternoon in the school library separating earthworms, preparing habitats, and treating incisions with silver.

Applying colloidal silver to the wound on the earthworm ….

Measuring regrowth from day to day

We had to get creative in finding the wounds and measuring them. Under a black light, the pyranine made the silver easier to see and measure.

InVivoExperimentation - Test1- MeasurementofIncision(Initial=1.0cm)

ConditionApplied Day3 Day4 Day5

Earthworm1 Earthworm2 Earthworm3 Earthworm1 Earthworm2 Earthworm3 Earthworm1 Earthworm2 Earthworm3

ControlNoSilver 1 1 0.8 0.7 0.8 0.8 0.5 0.5 0.4

ColloidalSilver 0.7 0.7 0.6 0.3 0.4 0.5 0 0.1 0.2

0

0.2

0.4

0.6

0.8

1

1.2

1 2 3


0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

1 2 3


Measurement 1, 2, 3Measurement 1, 2, 3

Leng

th o

f Inc

ision

cm

Leng

th o

f Inc

ision

cm

InVivoExperimentation - Test2- MeasurementofIncision(Initial=1.0cm)



ControlNoSilver 0.9 0.8 1 0.7 0.5 0.7 0.5 0.4 0.5

ColloidalSilver 0.8 0.7 0.6 0.4 0.3 0.2 0 0 0

0

0.2

0.4

0.6

0.8

1

1 2 3


00.10.20.30.40.50.60.70.8

1 2 3


Measurement 1, 2, 3

Leng

th o

f Inc

ision

cm

Measurement 1, 2, 3

Leng

th o

f Inc

ision

cm

InVivoExperimentation - Test3- MeasurementofIncisionInitial=1.0cm



ControlNoSilver 0.9 1 0.7 0.8 0.8 0.7 0.6 0.4 0.5

ColloidalSilver 0.6 0.7 0.5 0.3 0.4 0.2 0.1 0.2 0

00.10.20.30.40.50.60.70.8

1 2 3

Mean Result Test 3: Control No Silver

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

1 2 3

Mean Result Test 3: Colloidal Silver

Measurement 1, 2, 3

Leng

th o

f Inc

ision

cm

Measurement 1, 2, 3

Leng

th o

f Inc

ision

cm

During tests conducted in vivo using the earthworm Lubricus terrestris for testing, a 1 cm incision was made into the skin of the worm just below the clitellum band. Silver was mixed with pyranine, a non-toxic chemical that glows brilliantly under UV black light. Under a black light, it was easy to see the site of the incision and makemeasurements, whereas it was extremely difficult before this treatment.

Then, the silver was applied to the wound. Earthworms were returned to their containers and measurements were made in cm on Day 3, 4, and 5. The worms treated with silver had greater cell growth and wound healing faster than those with no silver applied. These results were consistent during testing and show

that colloidal silver does speed cell regrowth and healing on living organisms.

Bandage Enhancement

´ Photos * Data * Tables * Graphs

´ Silver Bullet Ecybermission Team 2016

Embedding Hydrogel with Silver

Trial AmountofSilver inSyringe(ml) PercentofSilverSuccessfullyTransferredtoHydrogel(ml)

1 0.5 100%

2 0.5 100%

3 0.5 85%

4 0.5 100%

5 0.5 100%

In every test except Test 3, 100% of the silver that was contained in the syringe was able to be successfully transferred into the hydrogel pad. The silver remained in the hydrogel as part of the bandage throughout testing. This hydrogel enhanced bandage with colloidal silver would keep a wound moist, protected from airborne bacteria, and in contact with colloidal silver during the time the bandage remains on the wound.

Adding silver from a syringe to a hydrogel pad to enhance the bandage’s ability to fight bacteria.

IN VITRO EXPERIMENTATION

Effective Materials in Preventing

Bacterial Growth and Halting Biofilms

Silver Bullet

eCYBERMISSION

7th Grade 2016

• To identify a material that will be highly effective in preventing bacterial growth for the purpose of engineering an ointment for chronic wounds

Purpose

Hypothesis

• Following extensive research, it is believed that if a composite material of silver + xylitol is applied in vitro, then bacterial growth will be stopped more effectively than with the application of silver, xylitol, or probiotics alone.

• During in vitro tests, if the zone of inhibition surrounding the applied materials (silver, xylitol, xylitol + silver, or fructooligosaccaride probiotic) is at least 10.0 mm in diameter, the hypothesis will be proved valid. This would indicate a clear area free of bacterial growth as a result of the material application.

• Measurements will be made using a metric ruler from the center of a material-soaked disk outward to the nearest bacterial colony.

• The disks are used to control the size of application of the material within the Petri dish for accurate and reliable measurements. According to Dr. Rumbaugh in the wound care laboratory, a zone at least double the size of application would indicate a successful treatment.

Validity of the Hypothesis

• Controls – A Petri dish containing agar and bacteria will be compared to the experimental Petri dishes

• Constants – To each dish in every trial, the temperature will be maintained at 37oC for incubation, the same amount of material will be applied to the filter paper, and the zone of inhibition around each sample will be read at the same time.

Controls and Constants

Manipulated Independent Variable

• The independent variable in this experiment is the type of material used in the Petri dish to stop the bacteria.

• Probiotics Colloidal Silver Xylitol Silver + Xylitol

Responding Dependent Variable • The dependent variable is the measurement taken called the Zone of

Inhibition. The area surrounding the material disk and which is free from bacteria will be measured in millimeters. This is a measure of how effective the material is in stopping the growth of bacteria

1. Measure 23 grams of nutrient agar using a triple beam balance.

2. Measure one liter of distilled water into a saucepan and add the 23 grams of agar to the pan.

3. Heat and stir until the nutrient agar dissolves and then boil for one minute.

4. Carefully pour the solution into sterile Petri dishes, being sure to completely cover the bottom of the Petri dish.

5. Place lids on the Petri dishes, turn them upside down to prevent condensation from collecting on the agar, and allow to cool to 50oC and until the gel is set.

6. Follow Dr. Rumbaugh’s lab directions and touch the agar gently to inoculate it with bacteria that would commonly be found near chronic wounds. Cough on the agar dish one time as well to simulate the bacteria found in the air that could cause infection in wounds.

7. Place paper disks saturated in xylitol, colloidal silver, probiotics, and silver + xylitol into four divided sections of the Petri dishes.

8. Incubate at 37oC or body temperature for 16 hours.

9. Remove Petri dishes from incubator and measure (mm) the Zone of Inhibition from the edge of the disk to the area clear of bacteria surrounding the disk.

Procedure for In Vitro Experimentation

Zone of Inhibition Measurements

The black represents the

bacteria and the blue

spots represent the

saturated paper disk of

colloidal silver, xylitol,

probiotics, or silver +

xylitol. The ring around

the material disk is clear

of bacteria on some

samples and bacteria

has overtaken the disk in

others. The most effective

materials are those that

prevent bacteria from

getting near.

In Vitro Experimentation: Trial 1 Testing the Effectiveness of Materials in Preventing Bacterial Growth In Vitro

Trial 1

Material Sample 1 Sample 2 Sample 3 Sample 4 Mean


Probiotics 9 8 6 3 6.5

Xylitol 4 12 5 3 6

Silver 15 15 5 10 11.25

Composite - Silver + Xylitol 6 12 4 5 6.8

Control 5 3 2 8 4.5

0 2 4 6 8 10 12

Probiotics

Xylitol

Silver


Control

Effective Materials for Preventing Bacterial Growth:

Trial 1


In Vitro Experimentation: Trial 2

Trial 2



Probiotics 6 8 5 5 6

Xylitol 4 11 6 7 7

Silver 20 25 15 20 20

Composite - Silver + Xylitol 3 12 7 13 8.8

Control 5 3 2 8 4.5

0 5 10 15 20 25

Probiotics

Xylitol

Silver


Control

Effective Materials for Preventing Bacterial Growth:

Trial 2


In Vitro Experimentation: Trial 3

Trial 3



Probiotics 7 7 10 2 6.5

Xylitol 2 7 9 8 6.5

Silver 24 23 19 22 17.6

Composite - Silver + Xylitol 1 9 6 12 7

Control 2 3 8 3 4

0 2 4 6 8 10 12 14 16 18 20

Probiotics

Xylitol

Silver


Control

Effective Materials for Preventing Bacterial

Growth: Trial 3


Qualitative Data/Observations/Results

Silver was the most effective ingredient tested in vitro for

stopping bacterial growth. In Trial 1, Trial 2, and Trial 3

consistent results were obtained in sample after sample tested.

Our hypothesis was refuted because we expected the

composite of silver + xylitol to "outperform" silver treatments

alone. Based on the results of repeated testing, silver will now

be tested in vivo to learn if similar success in fighting bacteria

and promoting cell regrowth/healing will occur on a living

organism. Testing will NOT involve vertebrates - invertebrates

only will be used for experimentation.

• The most effective material tested in fighting biofilms and bacteria that cause infection was colloidal silver. Our hypothesis was refuted – the composite of silver + xylitol was ineffective, while silver alone consistently showed great potential in fighting bacteria.

• With the knowledge of silver and its effectiveness in stopping bacteria, the next stage of testing will involve the invertebrate earthworms and in vivo experimentation.

Conclusion

• In vitro testing

• Identification of silver as antimicrobial

Step 1

In Vitro

• In vivo testing

• Silver used on earthworms with cuts

Step 2

In Vivo • Engineer an ointment

• Silver used in hydrogels

Step 3

Ointment

Progression from In Vitro Testing to the Development of a Salve for Chronic Wounds

IN VITRO EXPERIMENTATION

Effective Materials in Preventing

Bacterial Growth and Halting Biofilms

Silver Bullet eCYBERMISSION 7th Grade 2016

• To identify a material that will be highly effective in preventing bacterial growth for the purpose of engineering an ointment for chronic wounds

Purpose

Hypothesis

• Following extensive research, it is believed that if a composite material of silver + xylitol is applied in vitro, then bacterial growth will be stopped more effectively than with the application of silver, xylitol, or probiotics alone.

• During in vitro tests, if the zone of inhibition surrounding the applied materials (silver, xylitol, xylitol + silver, or fructooligosaccaride probiotic) is at least 10.0 mm in diameter, the hypothesis will be proved valid. This would indicate a clear area free of bacterial growth as a result of the material application.

• Measurements will be made using a metric ruler from the center of a material-soaked disk outward to the nearest bacterial colony.

• The disks are used to control the size of application of the material within the Petri dish for accurate and reliable measurements. According to Dr. Rumbaugh in the wound care laboratory, a zone at least double the size of application would indicate a successful treatment.

Validity of the Hypothesis

• Controls – A Petri dish containing agar and bacteria will be compared to the experimental Petri dishes

• Constants – To each dish in every trial, the temperature will be maintained at 37oC for incubation, the same amount of material will be applied to the filter paper, and the zone of inhibition around each sample will be read at the same time.

Controls and Constants

Manipulated Independent Variable • The independent variable in this experiment is the type of material

used in the Petri dish to stop the bacteria.

• Probiotics Colloidal Silver Xylitol Silver + Xylitol

Responding Dependent Variable • The dependent variable is the measurement taken called the Zone of

Inhibition. The area surrounding the material disk and which is free from bacteria will be measured in millimeters. This is a measure of how effective the material is in stopping the growth of bacteria

1. Measure 23 grams of nutrient agar using a triple beam balance.

2. Measure one liter of distilled water into a saucepan and add the 23 grams of agar to the pan.

3. Heat and stir until the nutrient agar dissolves and then boil for one minute.

4. Carefully pour the solution into sterile Petri dishes, being sure to completely cover the bottom of the Petri dish.

5. Place lids on the Petri dishes, turn them upside down to prevent condensation from collecting on the agar, and allow to cool to 50oC and until the gel is set.

6. Follow Dr. Rumbaugh’s lab directions and touch the agar gently to inoculate it with bacteria that would commonly be found near chronic wounds. Cough on the agar dish one time as well to simulate the bacteria found in the air that could cause infection in wounds.

7. Place paper disks saturated in xylitol, colloidal silver, probiotics, and silver + xylitol into four divided sections of the Petri dishes.

8. Incubate at 37oC or body temperature for 16 hours.

9. Remove Petri dishes from incubator and measure (mm) the Zone of Inhibition from the edge of the disk to the area clear of bacteria surrounding the disk.

Procedure for In Vitro Experimentation

Zone of Inhibition Measurements The black represents the bacteria and the blue spots represent the saturated paper disk of colloidal silver, xylitol, probiotics, or silver + xylitol. The ring around the material disk is clear of bacteria on some samples and bacteria has overtaken the disk in others. The most effective materials are those that prevent bacteria from getting near.

In Vitro Experimentation: Trial 1 Testing the Effectiveness of Materials in Preventing Bacterial Growth In Vitro

Trial 1


Size of Zone of Inhibition Surrounding the Disk (mm) Probiotics 9 8 6 3 6.5

Xylitol 4 12 5 3 6 Silver 15 15 5 10 11.25

Composite - Silver + Xylitol 6 12 4 5 6.8 Control 5 3 2 8 4.5

0 2 4 6 8 10 12

Probiotics

Xylitol

Silver


Control



In Vitro Experimentation: Trial 2 Trial 2


Size of Zone of Inhibition Surrounding the Disk (mm) Probiotics 6 8 5 5 6

Xylitol 4 11 6 7 7 Silver 20 25 15 20 20

Composite - Silver + Xylitol 3 12 7 13 8.8 Control 5 3 2 8 4.5

0 5 10 15 20 25

Probiotics

Xylitol

Silver


Control



In Vitro Experimentation: Trial 3 Trial 3


Size of Zone of Inhibition Surrounding the Disk (mm) Probiotics 7 7 10 2 6.5

Xylitol 2 7 9 8 6.5 Silver 24 23 19 22 17.6

Composite - Silver + Xylitol 1 9 6 12 7 Control 2 3 8 3 4

0 2 4 6 8 10 12 14 16 18 20

Probiotics

Xylitol

Silver


Control



Qualitative Data/Observations/Results

Silver was the most effective ingredient tested in vitro for stopping bacterial growth. In Trial 1, Trial 2, and Trial 3 consistent results were obtained in sample after sample tested. Our hypothesis was refuted because we expected the composite of silver + xylitol to "outperform" silver treatments alone. Based on the results of repeated testing, silver will now be tested in vivo to learn if similar success in fighting bacteria and promoting cell regrowth/healing will occur on a living organism. Testing will NOT involve vertebrates - invertebrates only will be used for experimentation.

• The most effective material tested in fighting biofilms and bacteria that cause infection was colloidal silver. Our hypothesis was refuted – the composite of silver + xylitol was ineffective, while silver alone consistently showed great potential in fighting bacteria.

• With the knowledge of silver and its effectiveness in stopping bacteria, the next stage of testing will involve the invertebrate earthworms and in vivo experimentation.

Conclusion

• In vitro testing • Identification

of silver as antimicrobial

Step 1 In Vitro

• In vivo testing • Silver used on

earthworms with cuts

Step 2 In Vivo • Engineer an

ointment • Silver used in

hydrogels

Step 3 Ointment

Progression from In Vitro Testing to the Development of a Salve for Chronic Wounds

SilverBulletWebsite

Theteamisdevelopingawebsitetospreadawarenessofchronicwoundsandthealternativemethodsoftreatmentthatmaybeavailabletostopinfection.ThisisthefirstwebsiteI’vebuiltandI’mlearningalotabouthowtoaddimportantfeaturesthatwillmeettheneedsofpeoplewithchronicwounds.Continuecheckingbacktoseetheprogressbeingmade.

http://silverbulletecm.weebly.com

QRCode

Tomakeiteasyforpeopletovisitourwebsite,theteamcreatedaQRcodetothewebsiteonpostersfortheplaceswewouldbepresentinginformation.Thisisalsoavailableonourbusinesscardsandinbrochures.

team advisors - national science teachers...

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